Methods for treating inflammation and pain

ABSTRACT

The present invention is directed to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and an ω-(methanesulfonyl)alkenylnitrile compound, or a pharmaceutically acceptable salt thereof. The present invention is also directed to a method for treating inflammation, inflammatory-related disorders, or pain, by administering an ω-(methanesulfonyl)alkenylnitrile compound or a pharmaceutically acceptable salt or solvate thereof to a subject in need thereof. Oral and topical administration are preferred.

This application is a continuation-in-part of PCT/US2013/059094, filedSep. 10, 2013, which claims the priority of U.S. Provisional ApplicationNo. 61/699,720, filed Sep. 11, 2012. This application also claims thebenefit of U.S. Provisional Application No. 61/955,683, filed Mar. 19,2014. The above-identified applications are incorporated herein byreference in their entireties.

FIELD OF THE INVENTION

The present invention relates to a pharmaceutical composition comprisinga pharmaceutically acceptable carrier and anω-(methanesulfonyl)alkenylnitrile compound, or its pharmaceuticallyacceptable salts. The present invention also relates to methods of usingthe compound for treating inflammation or inflammatory-related disordersand pain. Oral and topical routes of administration are preferred.

BACKGROUND OF THE INVENTION

Inflammation is a process by which microbes or tissue injury induce therelease of cytokines and chemokines from various cell types producingincreased blood vessel permeability, upregulation of endothelialreceptors, and thus increased egress of various cells of the innate andadaptive immune system which enter surrounding tissue and grosslyproduce the classical picture of inflammation, i.e. redness, swelling,heat and pain.

Inflammation is a localized reaction of live tissue due to an injury,which may be caused by various endogenous and exogenous factors. Theexogenous factors include physical, chemical, and biological factors.The endogenous factors include inflammatory mediators, antigens, andantibodies. Endogenous factors often develop under the influence of anexogenous damage. An inflammatory reaction is often followed by analtered structure and penetrability of the cellular membrane. Endogenousfactors, such as mediators and antigens define the nature and type of aninflammatory reaction, especially its course in the zone of injury. Inthe case where tissue damage is limited to the creation of mediators, anacute form of inflammation develops. If immunologic reactions are alsoinvolved in the process, through the interaction of antigens,antibodies, and autoantigens, a long-term inflammatory process willdevelop. Various exogenous agents, for example, infection, injury,radiation, also provide the course of inflammatory process on amolecular level by damaging cellular membranes which initiatebiochemical reactions.

Based on the physical causes, pain can be divided into three types:nociceptive, neuropathic, and mix-type.

Nociceptive pain is the term for pain that is detected by specializedsensory nerves called nociceptors. These nerves are located throughoutthe soft tissues, such as muscles and skin, as well as the internalorgans. There are two types of nociceptive pain: somatic pain andvisceral pain. Visceral pain comes from the internal organs. Deepsomatic pain is initiated by stimulation of nociceptors in ligaments,tendons, bones, blood vessels, fasciae and muscles, and is dull, aching,poorly localized pain. Examples include sprains and broken bones.Superficial pain is initiated by activation of nociceptors in the skinor other superficial tissue, and is sharp, well-defined and clearlylocated. Examples of injuries that produce superficial somatic paininclude minor wounds and minor (first degree) burns. Nociceptive pain isusually short in duration and end when the damage recovers. Examples ofnociceptive pain include postoperative pain, sprains, bone fractures,burns, bumps, bruises, and inflammatory pain.

Neuropathic pain is pain caused by damage or disease that affects thesomatosensory system. Neuropathic pain is originated from spontaneousectopic neuron discharge in the nervous system either in central or inperipheral. Because the underlying etiologies are usually irreversible,most neuropathic pain are chronic pain. Most people describe neuropathicpain as shooting, burning, tingling, lancinating, electric shockqualities, numbness, and persistent allodynia. The nomenclature ofneuropathic pain is based on the site of initiating nervous system withthe etiology; for examples, central post-stroke pain, diabetesperipheral neuropathy, post-herpetic (or post-shingles) neuralgia,terminal cancer pain, phantom limb pain.

Mix-type pain is featured by the coexistence of both nociceptive andneuropathic pain. For example, muscle pain trigger central or peripheralneuron sensitization leading to chronic low back pain, migraine, andmyofacial pain.

Connective tissues are subjected to a constant barrage of stress andinjury. Acute or chronic impacts and the natural progression of variousdegenerative diseases all produce painful inflammation in joint regions,such as the neck, back, arms, hips, ankles and feet. These afflictionsare common and often debilitating.

Current therapy is directed to some or all of the pathogeneticcomponents of inflammation. For example, corticosteroids have a broadspectrum of activities and NSAIDS are more specificallyanti-prostaglandin and analgesic. All current therapies have relativelyhigh rates of adverse effects and adverse effects are severe andserious.

There is a need for a composition and a method for treatinginflammation, inflammatory-related disorders, and pain. The compositionshould be economic and easy to manufacture, and the method should beeffective and have no significant side effects.

SUMMARY OF THE INVENTION

The present invention is directed to a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier and anω-(methanesulfonyl)alkenylnitrile compound or a pharmaceuticallyacceptable salt or solvate thereof. The compound is at least 90% pure(w/w).

The present invention is also directed to a method for treatinginflammation, inflammatory-related disorders, and pain. The methodcomprises the step of administering an ω-(methanesulfonyl)alkenylnitrilecompound or a pharmaceutically acceptable salt thereof to a subject inneed thereof. The pharmaceutical composition comprising the activecompound can be applied by any accepted mode of administration includingoral, topical, and parenteral (such as intravenous, intramuscular,subcutaneous or rectal). Oral and topical administration are preferred.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the latency in seconds of mouse tail flick when measured at0, 5, 15, 30, 45, 60, and 75 minutes post morphine dosing. The mice weretreated with vehicle, morphine alone, or morphine plus the activecompound 3-(methylsulfonyl)acrylonitrile. The active compound was dosedan hour before morphine dosing.

DETAILED DESCRIPTION OF THE INVENTION Definition

“Alkyl” refers to groups of from 1 to 12 carbon atoms, either straightchained or branched, preferably from 1 to 8 carbon atoms, and morepreferably 1 to 6 carbon atoms.

“Arylalkyl” refers to aryl -alkyl—groups preferably having from 1 to 6carbon atoms in the alkyl moiety and from 6 to 10 carbon atoms in thearyl moiety. Such arylalkyl groups are exemplified by benzyl, phenethyland the like.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 12 carbon atomshaving a single cyclic ring or multiple condensed rings which can beoptionally substituted with from 1 to 3 alkyl groups. Such cycloalkylgroups include, by way of example, single ring structures such ascyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl,2-methylcyclopentyl, 2-methylcyclooctyl, and the like, or multiple ringstructures such as adamantyl, and the like.

“Pharmaceutically acceptable salts”, as used herein, are salts thatretain the desired biological activity of the parent compound and do notimpart undesired toxicological effects.

Pharmaceutically acceptable salt forms include various crystallinepolymorphs as well as the amorphous form of the different salts. Thepharmaceutically acceptable salts can be formed with metal or organiccounterions and include, but are not limited to, alkali metal salts suchas sodium or potassium; alkaline earth metal salts such as magnesium orcalcium; and ammonium or tetraalkyl ammonium salts, i.e., NX₄+ (whereinX is C₁₋₄).

“Solvates,” as used herein, are addition complexes in which the compoundis combined with an acceptable co-solvent in some fixed proportion.Co-solvents include, but are not limited to, ethyl acetate, lauryllactate, myristyl lactate, cetyl lactate, isopropyl myristate, ethanol,1-propanol, isopropanol, 1-butanol, isobutanol, tert-butanol, acetone,methyl ethyl ketone, and diethyl ether.

The inventors have discovered that ω-(methanesulfonyl)alkenylnitrilesare effective for treating inflammation, inflammatory-related disorders,and pain.

ω-(Methanesulfonyl)alkenylnitriles

ω-(Methanesulfonyl)alkenylnitriles (methanesulfonylalkanenitriles)useful for the present invention are compounds of formula I (both E- andZ-isomers), or a pharmaceutically acceptably salt or solvate thereof:

wherein R₁ and R₂ are independently selected from the group consistingof H, straight-chain alkyl, branched alkyl, cycloalkyl, and arylalkyl,and halogen; and

n=0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; m=0, 1, 2, 3, 4, 5, 6, 7, 8, or 9;and n+m=0-9. Preferably n+m=0-4, more preferably n=0-4 and m=0.

The structures of (E)-methanesulfonylacrylonitrile (1; n=m=0; R₁═R₂═H;molecular weight MW=131.15), (Z)-methanesulfonylacrylonitrile (2; n=m=0;R₁═R₂═H; MW=131.15), 2, 3-dichloro-3-methanesulfonylacrylonitrile (3;n=m=0; R₁═R₂═Cl, MW=200.04), 4-methanesulfonylbut-2-enenitrile (4; n=1,m=0; R₁═R₂═H, MW=145.18), 5-methanesulfonylpent-2-enenitrile (5; n=2,m=0; R₁═R₂═H, MW=159.21), 6-methanesulfonylhex-2-enenitrile (6; n=3,m=0; R₁═R₂═H, MW=173.23), and 7-methanesulfonylhept-2-enenitrile (7;n=4, m=0; R₁═R₂═H, MW=187.26) are shown below.

(E)-Methanesulfonylacrylonitrile (1) can be prepared according to U.S.Pat. No. 3,541,119. For example, the compound can be prepared byalkylating sodium methanesulfinate with 2,3-dichloropropionitrile in thepresence of a base.

(Z)-Methanesulfonylacrylonitrile (2) can be prepared by modifying theprocedure of Asscher and Vofsi (J. Chem. Soc. 1964, 4962-4971). Forexample, the compound can be prepared by a copper-catalyzed addition ofmethanesulfonyl chloride to acrylonitrile.

2,3-dichloro-3-methanesulfonylacrylonitrile (3) can be preparedaccording to U.S. Pat. No. 4,424,167. For example, the compound can beprepared from 3-methylsulfonylpriopionitrile by a series ofchlorination/dehydrohalogenation reactions using chlorine gas in thepresence of a basic catalyst.

ω-Methanesulfonylalk-2-enenitriles (e.g. Compounds 4, 5, 6, and 7) canbe prepared by alkylating sodium methanesulfinate with requisiteω-halolalk-2-enenitriles, or by alkylating sodium thiomethoxide with therequisite ω-halolalk-2-enenitriles, followed by oxidation of theproducts with hydrogen peroxide or other appropriate oxidizing agentsuch as urea hydrogen peroxide.

Pharmaceutical Compositions

The present invention provides pharmaceutical compositions comprisingone or more pharmaceutically acceptable carriers and an active compoundof ω-(methanesulfonyl)alkenylnitrile, or a pharmaceutically acceptablesalt or a solvate thereof. The pharmaceutical composition can includeone of the cis or trans isomers, or both isomers either equimolar, or ofdifferent amounts. The active compound or its pharmaceuticallyacceptable salt or solvate in the pharmaceutical compositions in generalis in an amount of about 0.01-20%, or 0.05-20%, or 0.1-20%, or 0.2-15%,or 0.5-10%, or 1-5% (w/w) for a topical formulation; about 0.1-5% for aninjectable formulation, 0.1-5% for a patch formulation, about 1-90% fora tablet formulation, and 1-100% for a capsule formulation.

In one embodiment, the active compound is incorporated into anyacceptable carrier, including creams, gels, lotions or other types ofsuspensions that can stabilize the active compound and deliver it to theaffected area by topical applications. In another embodiment, thepharmaceutical composition can be in a dosage form such as tablets,capsules, granules, fine granules, powders, syrups, suppositories,injectable solutions, patches, or the like. The above pharmaceuticalcomposition can be prepared by conventional methods.

Pharmaceutically acceptable carriers, which are inactive ingredients,can be selected by those skilled in the art using conventional criteria.Pharmaceutically acceptable carriers include, but are not limited to,non-aqueous based solutions, suspensions, emulsions, microemulsions,micellar solutions, gels, and ointments. The pharmaceutically acceptablecarriers may also contain ingredients that include, but are not limitedto, saline and aqueous electrolyte solutions; ionic and nonionic osmoticagents such as sodium chloride, potassium chloride, glycerol, anddextrose; pH adjusters and buffers such as salts of hydroxide,phosphate, citrate, acetate, borate; and trolamine; antioxidants such assalts, acids and/or bases of bisulfite, sulfite, metabisulfite,thiosulfite, ascorbic acid, acetyl cysteine, cysteine, glutathione,butylated hydroxyanisole, butylated hydroxytoluene, tocopherols, andascorbyl palmitate; surfactants such as lecithin, phospholipids,including but not limited to phosphatidylcholine,phosphatidylethanolamine and phosphatidyl inositiol; poloxamers andpoloxamines, polysorbates such as polysorbate 80, polysorbate 60, andpolysorbate 20, polyethers such as polyethylene glycols andpolypropylene glycols; polyvinyls such as polyvinyl alcohol andpovidone; cellulose derivatives such as methylcellulose, hydroxypropylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose andhydroxypropyl methylcellulose and their salts; petroleum derivativessuch as mineral oil and white petrolatum; fats such as lanolin, peanutoil, palm oil, soybean oil; mono-, di-, and triglycerides; polymers ofacrylic acid such as carboxypolymethylene gel, and hydrophobicallymodified cross-linked acrylate copolymer; polysaccharides such asdextrans and glycosaminoglycans such as sodium hyaluronate. Suchpharmaceutically acceptable carriers may be preserved against bacterialcontamination using well-known preservatives, these include, but are notlimited to, benzalkonium chloride, ethylenediaminetetraacetic acid andits salts, benzethonium chloride, chlorhexidine, chlorobutanol,methylparaben, thimerosal, and phenylethyl alcohol, or may be formulatedas a non-preserved formulation for either single or multiple use.

For example, a tablet formulation or a capsule formulation of the activecompound may contain other excipients that have no bioactivity and noreaction with the active compound. Excipients of a tablet or a capsulemay include fillers, binders, lubricants and glidants, disintegrators,wetting agents, and release rate modifiers. Binders promote the adhesionof particles of the formulation and are important for a tabletformulation. Examples of excipients of a tablet or a capsule include,but not limited to, carboxymethylcellulose, cellulose, ethylcellulose,hydroxypropylmethylcellulose, methylcellulose, karaya gum, starch,tragacanth gum, gelatin, magnesium stearate, titanium dioxide ,poly(acrylic acid), and polyvinylpyrrolidone. For example, a tabletformulation may contain inactive ingredients such as colloidal silicondioxide, crospovidone, hypromellose, magnesium stearate,microcrystalline cellulose, polyethylene glycol, sodium starchglycolate, and/or titanium dioxide. A capsule formulation may containinactive ingredients such as gelatin, magnesium stearate, and/ortitanium dioxide.

For example, a patch formulation of the active compound may comprisesome inactive ingredients such as 1,3-butylene glycol, dihydroxyaluminumaminoacetate, disodium edetate, D-sorbitol, gelatin, kaolin,methylparaben, polysorbate 80, povidone, propylene glycol,propylparaben, sodium carboxymethylcellulose, sodium polyacrylate,tartaric acid, titanium dioxide, and purified water. A patch formulationmay also contain skin permeability enhancer such as lactate esters(e.g., lauryl lactate) or diethylene glycol monoethyl ether.

Topical formulations including the active compound can be in a form ofgel, cream, lotion, liquid, emulsion, ointment, spray, solution, andsuspension. The inactive ingredients in the topical formulations forexample include, but not limited to, lauryl lactate(emollient/permeation enhancer), diethylene glycol monoethyl ether(emollient/permeation enhancer), DMSO (solubility enhancer), siliconeelastomer (rheology/texture modifier), caprylic/capric triglyceride,(emollient), octisalate, (emollient/UV filter), silicone fluid(emollient/diluent), squalene (emollient), sunflower oil (emollient),and silicone dioxide (thickening agent).

In one embodiment, lauryl lactate (for example, at about 0.1-10%, orabout 0.2-5%, or about 0.5-5%) is included in the topical gelformulation. Lauryl lactate is considered safe for topicaladministration. Lauryl lactate is qualified for human use withinpharmaceutical and cosmetic products. Lauryl lactate when used in atopical formulation enhances the permeability of the compound.Preferably lauryl lactate is purified to achieve ≧90%, preferably ≧95%purity; the high purity mitigates the presence of hydrolytic andoxidative agents. In addition, DMSO at 0.1-20%, or 0.5-10% (w/w) in theformulation provides suitable solubility of the active compound.

In another embodiment, diethylene glycol monoethyl ether is included inthe topical gel formulation.

Method of Use

Inflammation is a process and a state of tissue pathology resulting fromactivation and continuation of activity of the innate and acquiredcomponents of the immune system. The arachidonic acid cascade andcytokine production and action in cell to cell interactions are criticalcomponents of immune activation and response, which lead toinflammation. Arachidonic acid is a component of membrane phospholipids.After it is freed from phospholipids, arachidonic acid acts as aprecursor to many of the known eicosanoids including prostaglandins andleucotrienes, which are known pro-inflammatory entities.

The active compound is effective in inhibiting pro-inflammatory cytokinerelease (e.g., IL-1β, IL-6, TNFα, IL-4 and IFNγ) from human peripheralblood mononuclear cells in vitro. The active compound isanti-inflammatory when applied topically in the mouse ear swellingmodel, in which the inflammation is induced by arachidonic acid.

The present invention is directed to a method of treating inflammationand/or pain. ω-(Methanesulfonyl)alkenylnitrile, can be used as is, or itcan be administered in the form of a pharmaceutical composition thatadditionally contains a pharmaceutically acceptable carrier. The methodcomprises the steps of first identifying a subject suffering frominflammation and/or pain, and administering to the subject the activecompound, in an amount effective to treat inflammation and/or pain. “Aneffective amount,” as used herein, is the amount effective to treat adisease by ameliorating the pathological condition or reducing thesymptoms of the disease.

In one embodiment, the method reduces or alleviates the symptomsassociated with inflammation. The present invention provides a method totreat localized manifestation of inflammation characterized by acute orchronic swelling, pain, redness, increased temperature, or loss offunction in some cases.

In another embodiment, the present invention provides a method toalleviate the symptoms of pain regardless of the cause of the pain. Thegeneral term “pain” treatable by the present method includesnociceptive, neuropathic, and mix-type. The present invention reducespain of varying severity, i.e. mild, moderate and severe pain; acute andchronic pain. The present invention is effective in treating joint pain,muscle pain, tendon pain, burn pain, and pain caused by inflammationsuch as rheumatoid arthritis.

In one embodiment, the present invention is useful in treatinginflammation and/or pain associated in a musculoskeletal system. Thehighly innervated, musculoskeletal system has a high capacity fordemonstration of pain. In addition, the musculoskeletal system has ahigh capacity for tissue swelling. In musculoskeletal system, the degreeof tissue damage is frequently magnified out of proportion to theresulting inflammatory response.

The present invention provides a method for treating inflammation and/orpain associated with inflammatory skeletal or muscular diseases orconditions. The method comprises the steps of identifying a subject inneed thereof, and administering to the subject the active compound, inan amount effective to treat inflammation and/or pain. “The activecompound,” as used herein, is intended to include theω-(methanesulfonyl)alkenylnitrile compound (cis-isomer, trans-isomer, ora mixture thereof) and its pharmaceutically acceptable salts or solvatethereof. The skeletal or muscular diseases or conditions includemusculoskeletal sprains, musculoskeletal strains, tendonopathy,peripheral radiculopathy, osteoarthritis, joint degenerative disease,polymyalgia rheumatica, juvenile arthritis, gout, ankylosingspondylitis, psoriatic arthritis, systemic lupus erythematosus,costochondritis, tendonitis, bursitis, such as the common lateralepicondylitis (tennis elbow), medial epicondylitis (pitchers elbow) andtrochanteric bursitis, temporomandibular joint syndrome, andfibromyalgia.

In one embodiment, the present invention provides a method for treatinginflammation and/or pain associated with joints, ligaments, tendons,bone, muscles, or fascia. The method comprises the steps ofadministering to a subject in need thereof the active compound, in anamount effective to treat inflammation and/or pain.

The present invention also provides a method for treating pain by thecombined administration of morphine and methanesulfonylacrylonitrile.The method comprises the steps of: identifying a subject suffering frompain, and administering to the subject an effective amount of morphineand an effect amount of methanesulfonylacrylonitrile. The combinedtreatment can reduce the dosage of each single drug and can increase theanalgesic effects. In the combined treatement, morphine andmethanesulfonylacrylonitrile can be simultaneously or sequentiallyadministered. For simultaneous administration, morphine andmethanesulfonylacrylonitrile can be pre-mixed in one single formulationbefore administration. Alternatively, morphine andmethanesulfonylacrylonitrile can be separately administrated.

The present invention further provides a method for potentiating theanalgesic effect of morphine. The method comprises administering aneffect amount of ω-methanesulfonylacrylonitrile to a subject who suffersfrom pain and is being treated with morphine. The administration ofω-methanesulfonylacrylonitrile increases the analgesic effect ofmorphine, and thus less morphine can be used to achieve an optimalanalgesic effect. In addition, the analgesic effect is prolonged.

In one embodiment, the present invention is directed to a method oftreating inflammation and/or pain associated gout. Gout is a chronicinflammatory disease that is characterized by recurrent, sudden, andsevere attacks of acute inflammation (redness and tenderness) and painat the joints, often at the base of the big toe. Gout is caused byelevated levels of uric acid in the blood. Gout is a type of arthritis.Some people may develop chronic gout, which is also called goutyarthritis.

Skin is highly reactive to environmental stimuli and the epidermalcomponent of keratinocytes is a very rich source of both arachidonicacid and pro-inflammatory cytokines of IL-1 and TNF. The skin dendriticcells, Langerhans cells, recognize and process antigens for furtherimmune response of various lymphocytes and all of these cells areprimarily regulated by cytokines through their specific cell surfacereceptors.

ω-(methanesulfonyl)alkenylnitriles, which are effective in inhibitingarachidonic acid induced inflammation and in inhibiting the release ofpro-inflammatory cytokine, are effective to treat inflammation and/orpain associated with inflammatory skin diseases such as psoriasis, acne,rosacea, and dermatitis (e.g., contact dermatitis, and atopicdermatitis).

ω-(Methanesulfonyl)alkenylnitrile, which are effective in inhibitingarachidonic acid induced inflammation and in inhibiting the release ofpro-inflammatory cytokine, are effective to treat inflammatory skindiseases such as dermatitis (atopic dermatitis), psoriasis, acne, androsacea.

The present invention provides a method for treating inflammation and/orpain associated with inflammatory skin diseases such as psoriasis, acne,rosacea, and dermatitis, particularly contact dermatitis, and atopicdermatitis. The method comprises the steps of identifying a subject inneed thereof, and administering to the subject anω-(methanesulfonyl)alkenylnitrile, in an amount effective to treatinflammation and/or pain.

The present invention further provides a method for treatinginflammatory skin diseases such as dermatitis, psoriasis, and acne (Acnevulgaris). The method comprises the steps of identifying a subject inneed thereof, and administering to the subject anω-(methanesulfonyl)alkenylnitrile, in an amount effective to reduce oreliminate the symptoms of the disease.

Dermatitis (also called eczema) is generic inflammation of the skin.Specific types of dermatitis include atopic, contact, nummular, andphoto-induced.

Contact dermatitis is a localized rash or irritation of the skin causedby contact with a foreign substance. Only the superficial regions of theskin are affected in contact dermatitis. Inflammation of the affectedtissue is present in the epidermis (the outermost layer of skin) and theouter dermis (the layer beneath the epidermis). Contact dermatitisresults in large, burning, and itchy rashes. Contact dermatitis is aninflammatory condition of the skin either of irritant exposure to theskin without specific adaptive immunologic pathogenesis or of allergicsensitization and subsequent exposure of the skin to the sensitizingallergen with specific adaptive immunologic pathogenesis. Both involveinnate and acquired immune system response including arachidonic acidand cytokine components that initiate and propagate the disease throughcell to cell messaging by eicosanoid and/or cytokine moieties producedby epidermal cells, macrophages, dendritic cells, neutrophils,eosinophils, and various T and B lymphocytes. Contact dermatitis may beeither acute or chronic. The acute forms are pruritic with erythema,edema, and micro or macrovesiculation in the areas of skin contact bythe initiating factor. The chronic forms are pruritic with mildererythema, scaling, lichenification, and possibly fissuring particularlyon the hands.

Allergic contact dermatitis is a T cell-mediated delayed typehypersensitivity reaction that occurs upon hapten challenge insensitized individuals. The inflammatory response in classical allergiccontact dermatitis requires both a sensitization phase and anelicitation phase responsible for the recruitment and activation ofspecific T cells at the site of hapten skin challenge.

Atopic dermatitis is a genetically determined disease that is part ofthe broader disease complex of atopy that includes asthma, hay fever,and atopic dermatitis. Many individuals with atopic dermatitis havevarious mutations of the filaggrin gene that codes for an importantepidermal structural protein that when defective, results in abnormalbarrier function of the epidermis. The altered barrier allows exposureto multiple environmental allergens that are first recognized by innateimmune responses involving arachidonic acid and eicosanoids andrecruitment of eosinophils, mast cells, and other inflammatory cellsthat initiate an acute responses of itch, erythema, and subsequentscratching and additionally activate the adaptive immune responses thatinvolve inflammation by lymphocytes predominantly of a TH 2 derivationand activity. Atopic dermatitis is responsive to a number of cytokineinhibitors such as cyclosporine, and tacrolimus.

Current theory of the pathogenesis of psoriasis is that in individualswho are genetically susceptible a triggering event in the epidermis suchas injury or super antigen contact initiates an response of the innateimmune system with arachidonic acid and eicosanoid generation,recruitment and activity of neutrophils. Subsequent transformation ofthe response to that of a TH 1 adaptive immunity with cytokineactivation and activity of specific T lymphocytes effect thepathological changes in the epidermis and dermis, which result in thetypical psoriasis lesions of plaques that are erythematous, thickened,and scaly. Psoriasis is responsive to various immunomodulators includingcyclosporine, methotrexate, and a host of specific biologicals thatinterfere with cytokine signaling.

Acne vulgaris, a progressively inflammatory disorder of thepilosebaceous follicular unit especially of the face and upper chest andback is a very common disease of both males and females after initiationof puberty, and in females even prior to adrenal gland maturity.Increased production of androgenic hormones by adrenal, ovarian, andtesticular glands and by the pilosebaceous unit itself produce anincrease in sebum and changes in its lipid composition, which combinewith follicular epithelial cells to produce some degree of obstructionof the infra-infundibular portion of the pilosebaceous follicleresulting in the initial lesion of acne, the microcomedo. Thisconsequent dilation of the pore and the changed composition of sebum atpuberty facilitate colonization of the follicle by Propionibacteriumacnes bacilli that produce enzymes to degrade the triglycerides in sebumto free fatty acids that leak through the follicle into the dermis andincite arachidonic acid pathways of eicosanoid production and subsequentinitiation of inflammation. The bacilli also initiate chemokineproduction that attracts further inflammatory cells to the area andconsequent cytokine production and action to continue and amplifyinflammation. Thus initiation and propagation of progressiveinflammation in the microcomedo produces the evolution to the severalhallmark lesions of inflammatory acne, papule, pustule, nodule, andcyst. The present invention is useful to treat common acne, comedonicacne, papulopustular acne, papulocomedonic acne, nodulocystic acne, acneconglobata, cheloid acne of the nape of the neck, recurrent miliaryacne, necrotic acne, neonatal acne, occupational acne, acne rosacea,senile acne, solar acne or acne medicamentosa.

Rosacea is a chronic condition characterized by facial erythema andsometimes pimples. Rosacea typically begins as redness on the centralface across the cheeks, nose, or forehead, but can also less commonlyaffect the neck, chest, ears, and scalp. In some cases, additionalsymptoms, such as semi-permanent redness, telangiectasia (dilation ofsuperficial blood vessels on the face), red domed papules (small bumps)and pustules, red gritty eyes, burning and stinging sensations, and insome advanced cases, a red lobulated nose (rhinophyma), may develop.There are 3 subtypes of rosacea that affect the skin:erythematotelangiectatic rosacea, papulopustular rosacea, and phymatousrosacea. ω-(Methanesulfonyl)alkenylnitriles are effective in treatingatopic dermatitis and alleviating one or more symptoms selected from thegroup consisting of erythema, induration, lichenification, scaling, andoozing and crusting.

ω-(Methanesulfonyl)alkenylnitriles are effective in treating psoriasisand alleviating erythema, scaling, and/or thickness of the psoriasislesions.

ω-(Methanesulfonyl)alkenylnitriles are effective in treating acne andalleviating acne lesions selected from the groups consisting of closedcomedones, papules, pustules, nodules, and cysts.

ω-(Methanesulfonyl)alkenylnitriles are effective in treating rosacea andalleviating one or more symptoms selected from the group consisting oferythema, telangiectasia, red domed papules and pustules, red grittyeyes, and burning and stinging sensations.

The pharmaceutical composition of the present invention can be appliedby local administration and systemic administration. Localadministration includes topical administration. Systemic administrationincludes oral, parenteral (such as intravenous, intramuscular,subcutaneous or rectal), and other systemic routes of administration. Insystemic administration, the active compound first reaches plasma andthen distributes into target tissues. Topical administration and oraladministration are preferred routes of administration for the presentinvention.

Dosing of the composition can vary based on the extent of the injury andeach patient's individual response. For systemic administration, plasmaconcentrations of the active compound delivered can vary; but aregenerally 1×10⁻¹⁰-1×10⁻⁴ moles/liter, and preferably 1×10⁻⁸-1×10⁻⁵moles/liter.

In one embodiment, the composition is applied topically onto theaffected area and rubbed into it. The composition is topically appliedat least 1 or 2 times a day, or 3 to 4 times per day, depending on themedical issue and the disease pathology being chronic or acute. Ingeneral, the topical composition comprises about 0.01-20%, or 0.05-20%,or 0.1-20%, or 0.2-15%, 0.5-10, or 1-5% (w/w) of the active compound.For example, the topical composition comprises about 1 or 5% (w/w) ofthe active compound. Depending on the size of the affected area, 0.2-85mL, typically 0.2-10 mL, of the topical composition is applied to theindividual per dose. The active compound passes through skin and isdelivered to the site of discomfort.

In one embodiment, the pharmaceutical composition is administratedorally to the subject. The dosage for oral administration is generally0.1-20 mg/kg/day. For example, the dosage for oral administration is0.1-10, 0.5-10, or 1-10 mg/kg/day, for a human subject. For example, thedosage for oral administration is 1-600 mg/day, and preferably 1-100,10-50 mg/day for a human subject.

In one embodiment, the pharmaceutical composition is administratedintravenously to the subject. The dosage for intravenous bolus injectionor intravenous infusion is generally 0.03 to 5 or 0.03 to 1 mg/kg/day.

In one embodiment, the pharmaceutical composition is administratedsubcutaneously to the subject. The dosage for subcutaneousadministration is generally 0.3-20, 0.3-3, or 0.1-1 mg/kg/day.

Those of skill in the art will recognize that a wide variety of deliverymechanisms are also suitable for the present invention.

The present invention is useful in treating a mammal subject, such ashumans, horses, and dogs. The present invention is particularly usefulin treating humans.

The following examples further illustrate the present invention. Theseexamples are intended merely to be illustrative of the present inventionand are not to be construed as being limiting.

EXAMPLES Example 1 Preparation of 3-(methylsulfonyl)acrylonitrile

A methanolic solution of 2,3-dichloropropionitrile (60 mmol, a) wasadded dropwise to a mixture of sodium methanesulfinate (1 equivalent) inaqueous methanol containing sodium acetate (2 equivalent). The reactionwas monitored by TLC. After being stirred for about 1.5 hours, thereaction mixture was diluted with water and extracted with CHCl₃. TheCHCl₃ extract was washed with water, treated with charcoal andconcentrated. The residue was crystallized from hot ethanol. The yieldof 3-(methylsulfonyl)acrylonitrile (b) was 1.12 g (14%).

The product had a white, crystalline appearance. The melting point was106.5-107.5° C. The results of FTIR-ATR, ¹H NMR, ¹³C NMR analyses wereconsistent the structure of b. The elemental analysis found C 36.57, H3.84, and N 10.70. The GC-MS study showed m/z of 132.

Example 2 Gel Formulation 1

Table 1 exemplifies one gel formulation containingω-(methanesulfonyl)alkenylnitriles such as3-(methylsulfonyl)acrylonitrile.

TABLE 1 1% Gel Active compound 1.0% Dow Corning Elastomer Blend EL-69.0%  8050 ID Labrafac Lipophile WL 1349 8.0% Octisalate 5.0% LaurylLactate 3.2% Dimethyl Sulfoxide (DMSO) 1.8% Dow Corning 556 CosmeticGrade 7.9% Fluid Squalene 2.0% Sunflower Seed Oil 2.0% Dow CorningAerogel VM-2270 0.0% Total 100.0% 

Example 3 Gel Formulation 2

Table 2 exemplifies another gel formulation containingω-(methanesulfonyl)alkenylnitriles such as3-(methylsulfonyl)acrylonitrile.

TABLE 2 1-5% Gel Active compound 1.0-5.0% Diethylene glycol monoethylether  5.0% Acrylates/C10-30 alkyl acrylate crosspolymer (CARBOPOL ®Ultrez 20 0.50% polymer) Trolamine (tris(2-hydroxyethyl)amine) 0.47%Purified Water 89.03-93.03% Total 100.0% 

Example 4 In Vitro Inhibition of Cytokine Activities by3-(methylsulfonyl)acrylonitrile Objective

Test 3-(methylsulfonyl)acrylonitrile for inhibition of cytokinesecretion in peripheral blood mononuclear cells (PBMCs) stimulated withlipopolysaccharide (LPS).

-   Cytokines: IL-1β, IL-6, TNFα-   Test Compound Dose Concentrations: 50, 10, 5, 1, 0.5, 0.1 μM

Controls

-   Vehicle: DMSO, 0.1% v/v-   LPS control: LPS+Vehicle-   Cells negative control : Vehicle only/No LPS-   Positive control: Dexamethasone (100 nM, n=2)

Protocols

-   1. Microtiter plates were seeded with 1×10⁴ PBMCs/well in 150 μL    culture medium (RPMI 1640, 10% FBS, 1% Pen/Strep, 2 mM    L-Alanyl-L-glutamine)-   2. The plates were incubated at 37° C. in 5% CO₂ for 1 hour.-   3. 10 μL of test compound (at different concentrations),    dexamethasone, and vehicle controls were added to appropriate wells,    and the plates were incubated at 37° C. in 5% CO₂ for 1 hour.-   4. 40 μL of LPS (50 pg/mL final) were added and the plates were    incubated at 37° C. in 5% CO₂ for 1 hour.-   5. The plates were centrifuged at 1000 rpm for 10 minutes and    supernatants were collected.-   6. The supernatants were assayed for IL-1β, IL-6, TNFα levels using    the Luminex Bead kit.-   7. EC50 values were determined.

Results

The EC50's of 3-(methylsulfonyl)acrylonitrile for inhibiting IL-1β,IL-6, and TNFα were 0.16 μM 0.19 μM and 0.30 μM respectively. Theresults demonstrate that the active compound has an inhibitory effect oncytokines involved in the inflammatory process.

Example 5 Anti-inflammatory Activity of Oral Administration of3-(methylsulfonyl)acrylonitrile in Mice

3-(Methylsulfonyl)acrylonitrile was suspended in vehicle (1% Tween 80 inwater) to 0.1-2 mg/mL. The test compound, dexamethasone (positivecontrol), and vehicle were orally administered to mice and evaluated foranti-inflammatory activity in the topical arachidonic acid induced earswelling model in mice.

Male ICR derived mice weighing 22±2 g were used in this experiment. 5mice were used for each group (active compound, positive control, andvehicle). All animals were maintained in a controlled temperature(22-24° C.) and humidity (60%-70%) environment with 12-hour light/darkcycles for at least one week prior to use.

Arachidonic acid (0.5 mg in 20 μL acetone) was applied topically ontothe anterior and posterior surfaces of the right ear of test animals toinduce inflammation. Active compound of different concentrations invehicle (10 mL/kg) and vehicle (10 mL/kg) were orally administered bygavage 1 hour before arachidonic acid, whereas dexamethasone (0.3 mg/kg)was orally administered by gavage 3 hour before arachidonic acidchallenge. At 60 minutes after arachidonic acid induction of ear edema,the thickness of the right ear and the left ear was measured and thedifference calculated as an indication of the inflammation in the rightear. Significant activity was defined as a statistically significantinhibition (p-value determined by t-test was <0.05) in arachidonic acidinduced ear swelling relative to the vehicle-treated group. The resultsare shown in Table 3. Oral administration of3-(methylsulfonyl)acrylonitrile at dosage of 10 mg/kg caused significantinhibition in arachidonic acid induced ear swelling at 60 minutes.

TABLE 3 Dosage % Test Substance mg/kg Inhibition P Value Vehicle (1%Tween 80 NA NA in water) Dexamethasone 0.3 39 <0.005 (Positive control)Test Compound 0.1 11 0.364 Test Compound 0.3 11 0.364 Test Compound 1 90.397 Test Compound 3 13 0.172 Test Compound 10 26 0.042

Example 6 Potentiation of Analgesic Activity of Morphine by3-(methylsulfonyl)acrylonitrile in Mice Nociceptive Assays

The response of the mice to nociceptive stimuli was evaluated using thetime of tail-flick or tail-flick latency from 52° C. water bath.Briefly, the animal was placed in a restrainer with its tail outside.The tail (distal ⅓rd) was immersed into 52° C. water and the time forthe mouse to flick its tail out of the water was recorded. The maximumexposure permitted was 10 seconds. If there was no response within 10seconds, the animal tail was removed from the water to prevent damage tothe tail. Baseline data were collected using the tail flick procedure 2times over 1 hour, prior to any drug administration. The last baselineresponse data point was used as basal tail flick latency.

Drug Treatments

3-(methylsulfonyl)acrylonitrile was dissolved in 0.5% methyl cellulose(MC) to 2 mg/mL, and orally administered to mice (n=5) in a volume of 10ml/kg (20 mg/kg), 1 hour prior to morphine dosing. One mL plasticdisposal tuberculin syringes (Becton Dickinson & Co. Franklin Lakes,N.J.) and 20 gauge×38 mm flexible plastic feeding tubes (InstechLaboratories, Plymouth Meeting, Pa.) were used for oral drugadministration. Morphine at 5 mg/kg in saline was subcutaneouslyadministered to mice at time zero.

Tail flick measurements were made at 5, 15, 30, 45, 60, and 75 minutespost-morphine administration.

Data Analysis

Statistical evaluation was done using an appropriate analysis ofvariance (ANOVA) using tail flick latencies. Results of tail flickresponse from each group are calculated as mean+/−standard error ofmean. Analysis with p-values <0.05 is considered significant.

Results

The results are shown in FIG. 1. Comparing the latency of mice tailflick between morphine alone and morphine plus3-methanesulfonylacrylonitrile , the administration of3-methanesulfonylacrylonitrile increases the latency of tail flicksignificantly when measured at 30, 60, and 75 minutes post morphinedosing. The p values between morphine alone and morphine plusmethanesulfonylacrylonitrile at 30, 60, and 75 minutes post morphinedosing are 0.0033, 0.473, and 0.0045, respectively. Further, the resultsshow that morphine alone had no effect at 75 minutes post dosing,whereas morphine plus methanesulfonylacrylonitrile still had a stronganalgesic effects at 75 minutes post dosing.

Example 7 Anti-inflammatory and Analgesic Activity of the ActiveCompound by Oral Administration in a Carrageenan Model (PropheticExample)

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are suspended in vehicle (1% Tween 80 inwater) to 0.5-2 mg/mL. Active compounds, indomethacin (positivecontrol), and vehicle (1% Tween 80 in water) are orally administered andevaluated for anti-inflammatory and analgesic activity in the ratcarrageenan-induced paw inflammation model.

Rats are used in the experiment. Carrageenan (0.1 mL of a 1% suspension)is injected subcutaneously into the left hind paw to induceinflammation. Active compounds in vehicle (10 mL/kg) and vehicle areorally administered 1-2 hours before the carrageenan administration.Indomethacin is given orally at 5 mg/kg, 1 hour prior to carrageenanadministration. The degree of inflammation (edema, or swelling) isdetermined using a plethysmograph to measure paw volume. Analgesia isdetermined by measuring paw withdrawal to a mechanical stimulus usingvon Frey filaments. Inflammation and analgesia are measured 4 hoursafter carrageenan administration. ω-(methanesulfonyl)alkenylnitriles areexpected to have anti-inflammatory and/or analgesic properties asmeasured by a significant decrease in paw volume and/or a significantincrease in mechanical pressure needed to elicit paw withdrawal,respectively, as compared to the vehicle control.

Example 8 Anti-inflammatory and Analgesic Activity of Active Compound byTopical Administration in a Carrageenan Model (Prophetic Example)

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are prepared in the gel formulationaccording to Example 2 or 3.

Test materials in gel formulation (1-5%), indomethacin (positivecontrol), and vehicle (gel formulation without active compound), areevaluated for anti-inflammatory and analgesic activity in the ratcarrageenan-induced paw inflammation model.

Rats are used in the experiment. Carrageenan (0.1 mL of a 1% suspension)is injected subcutaneously into the left hind paw to induceinflammation. Test material (1-5%) or vehicle gel is applied to the pawtopically at volumes of 0.05, 0.1 0.15 or 2.0 mL, 1.5, 2.5, and 3.5hours following the carrageenan administration. Indomethacin is givenorally at 5 mg/kg, 1 hour prior to carrageenan administration. Thedegree of inflammation (edema, or swelling) is determined using aplethysmograph to measure paw volume. Analgesia is determined bymeasuring paw withdrawal to a mechanical stimulus using von Freyfilaments. Inflammation and analgesia are measured 4 hours aftercarrageenan administration. Test materials are expected to haveanti-inflammatory and/or analgesic properties as measured by asignificant decrease in paw volume and/or a significant increase inmechanical pressure needed to elicit paw withdrawal, respectively, ascompared to the vehicle control.

Example 9 Analgesic activity of the Active Compound by OralAdministration in a Hot Plate Model (Prophetic Example)

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are suspended in vehicle (1% Tween 80 inwater) to 0.5-2 mg/mL. Active compound, morphine (positive control), andvehicle (1% Tween 80 in water) are orally administered and evaluated foranalgesic activity in the rat hot plate model.

Rats are used in the experiment. Active compounds in vehicle (10 mL/kg)and vehicle (1% Tween 80 in water) are orally administered 1-2 hoursbefore the rat is placed on a 55° C. hot plate, and the time to lick thepaw is measured. The positive control, morphine, is given orally at 30mg/kg, 1 hour prior to hot plate testing.ω-(Methanesulfonyl)alkenylnitriles are expected to have analgesicproperties as measured by a significant increase in time to licking ascompared to the vehicle control (t-test, p<0.05).

Example 10 Analgesic activity of Active Compound in a Hot Plate Model byTopical Administration (Prophetic Example)

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are prepared in the gel formulationaccording to Example 2 or 3.

Test materials: active compound in gel formulation (1-5%), morphine(positive control), and vehicle (gel formulation without activecompound), are evaluated for analgesic activity in the rat hot platemodel.

Rats are used in the experiment. Test material (1-5%) or vehicle gel isapplied to the paw topically at volumes of 0.05, 0.1 0.15 or 2.0 mL. Onehour later the rat is placed on a 55° C. hot plate, and the time to lickthe paw is measured. The positive control, morphine, is given orally at30 mg/kg, 1 hour prior to hot plate testing. Test materials are expectedto have analgesic properties as measured by a significant increase intime to licking as compared to the vehicle control (t-test, p<0.05).

Example 11 Analgesic Activity of Active Compound in CFA-Induced ThermalHyperalgesia by Oral or Topical Administration (Prophetic Example)

CFA (Complete Freund's Adjuvant) is known to induce inflammatory pain.(Walker, et al. JPET. 304: 56-62, 2003. )

Male Sprague-Dawley rats weighing 180±20 g are used. The animals,divided into groups of 8-10each, receive a subplantar injection (0.1 ml)of CFA (0.1% solution) to the tested hindpaw at 24 hours prior toexperimentation. Thermal hyperalgesia is tested by using the IITCModel-336G (IITC INC.USA) apparatus with a thermally regulated glassfloors set at 30° C. Each rat is placed within a plastic box atop aglass floor. A light beam under the floor is aimed at the plantarsurface of the right hind paw. The time is measured automatically whenthe paw is withdrawn away from the thermal stimulus. The intensity ofthe light is adjusted with average group baseline latency from 12 to 14sec (pre-CFA) and a cut-off latency of 20 sec imposed. The latency towithdrawal is obtained for each rat and defined as the heat painthreshold. Twenty four hours after CFA injection, rats are pre-selected(with clear presence of thermal hyperalgesia) for experimentation onlyif the latency to withdrawal is less than 75% of baseline.

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are prepared in the gel formulationaccording to Example 2 or 3.

Active compound in gel formulation (1-5%), active compound in oralvehicle, morphine (positive control, p.o., 20 mg/kg), topical vehicle(gel formulation without an active compound), and oral vehicle (1% Tween80 in water) are evaluated for analgesic activity in the formalin model.

Test substance or vehicle is either administered orally (10-20 mg/kg) ortopically (1-5% gel formulation) to the plantar surface of the hind paw,at 60 minutes before the level of thermal hyperalgesia is again measured(post-treatment). Mean±SEM of thermal paw withdrawal time is calculated.Unpaired Student's t test is applied for comparison the values ofpost-treatment between test substance treated group and vehicle controlgroup. Positive activity is considered at P<0.05.

Example 12 Analgesic Activity of Active Compound in a Formalin Test byOral or Topical Administration (Prophetic Example)

Formalin test is a model of continuous pain resulting fromformalin-induced tissue injury. The formalin model encompassesinflammatory, neurogenic, and central mechanism of nociception. Theassay described below relates primarily to the late inflammatoryanalgesic phase sensitive to both strong central analgesic as well asweaker analgesic/anti-inflammatory agents (Hunskaar, et al., J.Neuroscience Meth. 14: 69-76, 1985). The formalin test represents asuitable model for testing compounds for treating neuropathic pain(Benson, et al. Proceedings of Measuring Behavior, 2008, Eds. Spink, etal, 324-325).

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are prepared in the gel formulationaccording to Example 3.

Active compound in gel formulation (1-5%), active compound in oralvehicle, morphine (positive control, p.o., 30 mg/kg), topical vehicle(gel formulation without an active compound), and oral vehicle (1% Tween80 in water) are evaluated for analgesic activity in the formalin model.

Test substance is administered to groups of 8-10 CD-1 derived male miceweighing 23±3 g one hour before subplantar injection of formalin (0.02ml, 2% solution). Test substance is either administered orally (10-20mg/kg) or topically (1-5% gel formulation) to the plantar surface of thehind paw. Reduction of the induced hind paw licking time recorded duringthe following 10 to 30 minute period by 50% or more indicates analgesicactivity. Positive activity is considered by a significant increase intime to licking in test compound as compared to the vehicle control(t-test, p<0.05).

Example 13 Analgesic Activity of Active Compound in Chronic ConstrictionInjury Model by Oral or Topical Administration (Prophetic Example)

Peripheral nerve lesions may generate a syndrome comprising, in additionto spontaneous pain, exaggerated responses to light touch (tactileallodynia). Chronic constriction injury model is a neuropathic painmodel.

Male Sprague Dawley rats weighing 180±20 g are used. Under pentobarbital(50 mg/kg, 5 ml/kg, i.p.) anesthesia, the sciatic nerve is exposed atmid-thigh level. Four ligatures (4-0 chromic gut), about 1 mm apart, areloosely tied around the nerve. The animals are then housed individuallyin cages with soft bedding for 7 days before testing. Constriction ofthe sciatic nerve produces nerve injury and unilateral neuropathic pain.

On the day of experiments, the animals have no access to food overnightbefore testing. The rats are placed under inverted plexiglass cages on awire mesh rack and allowed to acclimate for 20 to 30 minutes. Mechanicallodynia is evaluated by the Chaplan up/down method using von Freyfilaments to the plantar surface of the left hind paw. See Chaplan, etal. J. Neuroscience Methods, 53: 55-63, 1994.

Rats are pre-selected for experimentation only if the pain threshold7-14 days after nerve ligation (pre-treatment) is reduced by 10 grams offorce relative to the response of the individual paw before nerveligation (pre-ligation), namely, with clear presence of allodynia.

Active compounds of ω-(methanesulfonyl)alkenylnitrile such as3-(methylsulfonyl)acrylonitrile are prepared in the gel formulationaccording to Example 3.

Active compound in gel formulation (1-5%), active compound in oralvehicle, morphine (positive control, p.o., 20 mg/kg), topical vehicle(gel formulation without an active compound), and oral vehicle (1% Tween80 in water) are evaluated.

Test substance or vehicle is either administered orally (10-20 mg/kg) ortopically (1-5% gel formulation) to the plantar surface of the left hindpaw. The mechanical allodynia test is performed 30 min before(pre-treatment) and 1 and 3 hours after a single dose of test substanceor vehicle (post treatment). Paw withdraw thresholds of control andtested compound are measured.

Example 14 Treatment of Arthritis (Prophetic Example)

Zymosan injected directly into the knee joint of mice elicits aninflammatory response and is used as a model of arthritis (Verschure etal, Ann. Rheum Dis. 53:455-460, 1994). Endpoints measured in this modelinclude knee joint swelling score, cytokine levels in the synovialtissue and microscopic pathology of the knee.

Active compound 3-(methylsulfonyl)acrylonitrile (10-20 mg/kg in water,oral application) and vehicle control (water) are administered by oralgavage to mice with a volume of 5 mL/kg.

There are 5 mice per group, with a total of 10 knees injected. On Day 1,C57BL6mice are dosed (30 or 100 mg/kg/dose) with active compound orvehicle twice on Hours 0 and 12. On Day 2, mice are dosed with activecompound or vehicle on Hour 24, then injected intra-articularly with 180μg of zymosan (6 μL) into both knee joints on Hour 25, and then dosed asecond time on Hour 36 with each active compound or vehicle. On Day 3,mice are again dosed with active compound or vehicle on Hour 48. Twohour post-dosing on Hour 50, knees are scored for edema, synovial tissueis collected for measurement of cytokine levels, and knee joints areprocessed for histopathology for analysis of inflammatory immune cellinflux into the joint. Macroscopic joint swelling is assessed on allknees after the skin is removed using a scoring system ranging from 0 to3, with 0 being no swelling and 3 being severe swelling. Synovial tissueis taken from 5 knees for measurement of mouse interleukin-1β,interleukin-6, and interleukin-1 receptor antagonist levels. Theremaining 5 knees are processed for microscopic pathology for assessmentof cellular influx into the site of inflammation.

Results for each group are presented as mean±standard error of mean andstatistical evaluation is performed.

Treatment with active compound is expected to result in decreasedinflammation as measured by a decrease in joint swelling, decrease incytokine levels and decrease influx of inflammatory cells to the site ofinflammation.

Example 15 Treatment of Gout (Prophetic Example)

Monosodium urate monohydrate (MSU) crystals injected in combination witha free fatty acid (FFA) directly into the knee joint of mice elicits aninflammatory response and is used as a model of gout (Joosten et al,Arthritis & Rheumatism, 62(11):3237-3248, 2010)). Endpoints measured inthis model include knee joint swelling score, cytokine levels in thesynovial tissue and microscopic pathology of the knee.

Active compound 3-(methylsulfonyl)acrylonitrile (2-4 mg/mL in water,oral application) and vehicle control (water) are administered by oralgavage to mice with a volume of 5 mL/kg.

There are 5 mice per group, with a total of 10 knees injected. On Day 1,C57BL6 mice are dosed (50, 200, or 500 mg/kg/dose) with active compoundsor vehicle twice on Hours 0 and 12. On Day 2, mice are dosed with activecompounds or vehicle on Hour 24, then injected intra-articularly withMSU crystals (300 μg) and C18:0 FFA (200 μM, 10 μL) on Hour 25. Threehours later (Hour 28), knees are scored for edema, synovial tissue iscollected for measurement of cytokine levels, and knee joints areprocessed for histopathology for analysis of inflammatory immune cellinflux into the joint. Macroscopic joint swelling is assessed on allknees after the skin is removed using a scoring system ranging from 0 to3, with 0 being no swelling and 3 being severe swelling. Synovial tissueis taken from 5 knees for measurement of mouse interleukin-1f3,interleukin-6, and interleukin-1 receptor antagonist levels. Theremaining 5 knees are processed for microscopic pathology for assessmentof cellular influx into the site of inflammation.

Results for each group are presented as mean± standard error of mean andstatistical evaluation is performed.

Treatment with active compound is expected to result in decreasedinflammation as measured by a decrease in joint swelling, decrease incytokine levels and decrease influx of inflammatory cells to the site ofinflammation.

Example 16 Treatment of Knee Pain by Oral or Topical Administration(Prophetic Example)

Objectives: To investigate the efficacy of the active compound in a gelformulation or in an oral formulation in patients with mild to moderateknee pain associated with osteoarthritis following temporary cessationof standard NSAID therapy. The focus of this study is on the symptomscaused by painful arthritis. The clinical trial is utilizingosteoarthritis of the knee as a well-established paradigm for othermusculoskeletal disorders.

Topical Formulation: The gel formulation containing the active compound3-(methylsulfonyl)acrylonitrile at 1% and 5% (Example 3) are used inthis example. Placebo contains the same gel without the active compound.

Oral Formulation: Capsules or tablets each containing 50-200 mg of theactive compound 3-(methylsulfonyl)acrylonitrile are used in thisexample. Placebo capsules or tablets do not contain the active compound.

Methodology: A randomized, double-blind, placebo controlled, paralleltreatment multicenter clinical activity study.

Patients with painful osteoarthritis of the knee, controlled by a stabledose of standard NSAID therapy for at least 2 months, discontinue use ofthe NSAIDs for a 7-day washout period. Patients are then randomized in a1:1:1 ratio (1% active gel, 5% active gel, placebo). A total of up to150 patients are enrolled and treated for 14 days with follow-up at 14,21, and 28 days.

The active gel or placebo is applied to the affected knee 3 times a dayfor 14 days for a total of 42 treatments given every 4—6 hours whileawake.

The capsules or tablets are orally administered to patients 1-4 times aday for 14 days.

Patients are treated for 14 days and followed up for a further 14 days.NSAIDs may be restarted after the Day 14 visit.

Criteria for Evaluation:

Safety:

Adverse Events (AEs) throughout the study.

Physical examination at enrollment (−7 days, start of NSAID washoutperiod), Baseline, Day 14 and Day 28.

Vital signs at enrollment (−7 days, start of NSAID washout period),Baseline and Days 7, 14, 21, 28.

Clinical laboratory measurements at Baseline, and Days 7, 14, 21 and 28.

Clinical Activity:

The primary clinical activity parameters are the measurement of pain atthe site of application, as quantified by Pain on Movement assessment(100-mm VAS) and the Western Ontario and McMaster University (WOMAC)index (100-mm VAS or 5-point Likert scale). The effect of treatment onswelling, tenderness and inflammation of the knee is recorded, also thetime to reduction or eradication of pain after treatment is recorded.

Study Endpoints:

The primary clinical activity endpoints are:

Change from Baseline to Day 14 in WOMAC functional disability index andsub-indices:

Pain (Scale 0-20).

Stiffness (Scale 0-8).

Physical function (Scale 0-68).

Change from Baseline (Day 1) to Day 14 in Pain on Movement (1-100 mmVAS).

The secondary clinical activity endpoints are:

Change in Current Knee Pain score (100 mm VAS) at Baseline from pre-doseto 1 hour post-dose

Change in Current Knee Pain score (100 mm VAS) at Baseline from pre-doseto 2 hours post-dose

Change in Global Rating of Disease (5-point Likert scale)

Time to reduction or eradication of pain subsequent to each applicationof active compound.

Use of rescue medication (APAP).

Proportion of subjects experiencing an improvement in Pain on Movement(100-mm VAS) from Baseline to Day 14, equal to or greater than theminimum clinically important improvement (MCII) threshold of 15 mm or20%

-   -   Proportion of subjects whose Pain on Movement (100-mm VAS) at        Day 14 is less than the Patient Acceptable Symptom State (PASS)        threshold of 40 mm

Proportion of subjects who are ‘Responders’ based on the OARSI ResponderIndex, in relation to WOMAC Index.

Example 17 Treatment of Contact Dermatitis (Prophetic Example)

Mice dermally sensitized and challenged by dinitrofluorobenzene (DNFB)are used as a model of contact dermatitis (Saint-Mezard, J InvestDermatol, 120:641-647, 2003).

Sensitization and Challenging:

There are 5 mice per group. Each mouse is sensitized with 0.5% DNFB(vehicle=4:1 (vol/vol) acetone:olive oil) topically on the shavedabdomen, 6 days before challenge. The right ears of the mice are thenchallenged with a topical application of 0.2% DNFB in vehicle. The leftears of the mice receive the vehicle as control.

Oral Administration: Active compound 3-(methylsulfonyl)acrylonitrile(2-4 mg/mL in water) and control (water) are administered by oral gavageto mice with a volume of 5 mL/kg. Before challenge, each group of micereceive oral dosages of active compound or water at 24 hours, 12 hours,and 2 hours before the challenge.

After challenge, the same oral dosages of active compound or water aregiven to each mouse 7 hours, 22 hours, 31 hours, 46 hours, and 55 hoursafter the challenge. The thickness of the left and right ears aremeasured before challenge, and 24, 48, and 72 hours after challenge.Results are expressed as net swelling: thickness after challenge minusthickness before challenge. Net swelling of treated mice vs. controlmice are compared.

Topical Administration: Active compound(3-(methylsulfonyl)acrylonitrileprepared in vehicle (1:1;acetone:ethanol) at 375 mM and vehicle alone are topically applied toboth ears of the mice in a volume of 20 μl.

The topical doses are given after challenge to each mouse 7 hours, 22hours, 31 hours, 46 hours, and 55 hours after the challenge. Thethickness of the left and right ears are measured before challenge, and24, 48, and 72 hours after challenge. Results are expressed as netswelling: thickness after challenge minus thickness before challenge.Net swelling of treated mice vs. control mice are compared.

Example 18 Treatment of Atopic Dermatitis (Prophetic Example)

Objectives: To investigate the efficacy of3-(methylsulfonyl)acrylonitrile gel in patients having atopicdermatitis.

Topical Formulation: 3-(methylsulfonyl)acrylonitrile is prepared as agel formulation according to Example 3 or as a cream formulationaccording to Example 4. Active compounds in a gel or cream formulationare used in this experiment. Placebo contains the same gel or creamingredients without the active compound.

Oral Formulation: Capsules or tablets each containing 50-200 mg of theactive compound 3-(methylsulfonyl)acrylonitrile are used in thisexample. Placebo capsules or tablets do not contain the active compound.

Methodology: This is a randomized, double-blind, placebo controlled,parallel treatment clinical activity study.

Male and female patients with mild to severe atopic dermatitis areenrolled after discontinuation of all treatments for atopic dermatitisfor a period of 4 weeks before study initiation. Patients are randomizedin a 1:1 ratio (active gel, placebo). A total of 300 patients areenrolled and treated.

The active gel or placebo is applied twice a day to affected areas ofthe body for 12 weeks.

The capsules or tablets are orally administered to patients 1-4 times aday for 12 weeks.

The treatment results are evaluated at 2 week intervals until week 12and then at 4 weeks after discontinuation of the study medication.

Criteria for Evaluation:

Safety:

Safety is evaluated by general history and physical signs, laboratorytesting for hematology, serum chemistry, and urinalysis, and byevaluations of local application site tolerability parameters oferythema, scaling, dryness, stinging/burning utilizing a rating scale of“0” (None) to “3” (Severe).

Efficacy:

Efficacy is evaluated utilizing:

-   1. an overall assessment of disease severity at study entry and at 2    week intervals until week 12 and subsequently at 4 weeks after study    medication discontinuation. The investigator global assessment, IGA,    is based upon a rating scale of 0 to 4 with 0=none or clear,    1=almost clear, 2=mild disease involvement, 3=moderate disease    involvement, and 4=severe disease involvement, and:-   2. separate evaluation of a representative target atopic dermatitis    area of involvement for erythema, induration, lichenification,    scaling, and oozing and crusting with each parameter rated on a 0-4    scale with 0=none or clear, 1=almost clear, 2=mild disease    involvement, 3=moderate disease involvement, and 4=severe disease    involvement.

Statistical analyses of each of these efficacy evaluations are performedfor each of the 2 week study time points. Definitive evaluation ofefficacy is based upon comparisons of active to vehicle groups at end oftreatment at 12 weeks. The 4 week-post treatment evaluation is utilizedto evaluate durability of treatment effect after medicationdiscontinuation.

Example 19 Treatment of Psoriasis (Prophetic Example)

Objectives: To investigate the efficacy of the3-(methylsulfonyl)acrylonitrile gel in patients having psoriasisvulgaris.

Topical Formulation: 3-(methylsulfonyl)acrylonitrile is prepared as agel formulation according to Example 3 or as a cream formulationaccording to Example 4. Active compounds in a gel or cream formulationare used in this experiment. Placebo contains the same gel or creamingredients without the active compound.

Oral Formulation: Capsules or tablets each containing 50-200 mg of theactive compound 3-(methylsulfonyl)acrylonitrile are used in thisexample. Placebo capsules or tablets do not contain the active compound.

Methodology: This is a randomized, double-blind, placebo controlled,parallel treatment clinical activity study.

Male and female patients with mild to severe psoriasis vulgaris areenrolled. Patients discontinue all treatments for psoriasis for a periodof 4 weeks before study initiation. Patients are randomized in a 1:1ratio (active gel, placebo). A total of 200 patients are enrolled andtreated.

The active gel or placebo is applied twice a day to affected areas ofthe body for 12 weeks.

The capsules or tablets are orally administered to patients 1-4 times aday for 12 weeks.

The treatment results are evaluated at 2 week intervals until week 12and then at 4 weeks after discontinuation of the study medication.

Criteria for Evaluation:

Safety:

Safety is evaluated by general history and physical signs, laboratorytesting for hematology, serum chemistry, and urinalysis, and byevaluations of local application site tolerability parameters oferythema, scaling, dryness, stinging/burning utilizing a rating scale of“0” (None) to “3” (Severe).

Efficacy:

Efficacy is evaluated utilizing:

-   1. an overall assessment of disease severity at study entry and at 2    week intervals until week 12 and subsequently at 4 weeks after study    medication discontinuation. The investigator global assessment, IGA,    is based upon a rating scale of 0 to 4 with 0=none or clear,    1=almost clear, 2=mild disease involvement, 3=moderate disease    involvement, and 4=severe disease involvement, and:-   2. separate evaluation of a representative target psoriasis lesion    for erythema, scaling, and thickness of each parameter rated on a    0-4 scale with 0=none or clear, 1=almost clear, 2=mild disease    involvement, 3=moderate disease involvement, and 4=severe disease    involvement.

Statistical analyses of each of the efficacy evaluations are performedfor each of the 2 week study time points. Definitive evaluation ofefficacy is based upon comparisons of active to vehicle groups at end oftreatment at 12 weeks. The 4 week-post treatment evaluation is utilizedto evaluate durability of treatment effect after medicationdiscontinuation.

Example 20 Treatment of Acne (Prophetic Example)

Objectives: To investigate the efficacy of the3-(methylsulfonyl)acrylonitrile gel in patients having acne vulgaris.

Topical Formulation: 3-(methylsulfonyl)acrylonitrile is prepared as agel formulation according to Example 3 or as a cream formulationaccording to Example 4. Active compounds in a gel or cream formulationare used in this experiment. Placebo contains the same gel or creamingredients without the active compound.

Oral Formulation: Capsules or tablets each containing 50-200 mg of theactive compound (3-(methylsulfonyl)acrylonitrile are used in thisexample. Placebo capsules or tablets do not contain the active compound.

Methodology: This is a randomized, double-blind, placebo controlled,parallel treatment clinical activity study.

Male and female patients with mild to severe acne vulgaris are enrolled.Patients discontinue all treatments for acne for a period of 4 weeksbefore initiation of the study. Patients are randomized in a 1:1 ratio(active gel, placebo). A total of 500 patients are enrolled and treated.

The active gel or placebo is applied twice a day to affected areas ofthe body for 12 weeks.

The capsules or tablets are orally administered to patients 1-4 times aday for 12 weeks.

The treatment results are evaluated at 2 week intervals until week 12and then at 4 weeks after discontinuation of the study medication.

Criteria for Evaluation:

Safety:

Safety is evaluated by general history and physical signs, laboratorytesting for hematology, serum chemistry, and urinalysis, and byevaluations of local application site tolerability parameters oferythema, scaling, dryness, stinging/burning utilizing a rating scale of“0” (None) to “3” (Severe).

Efficacy:

Efficacy is evaluated utilizing:

-   1. an overall assessment of disease severity at study entry and at 2    week intervals until week 12 and subsequently at 4 weeks after    discontinuation of the study medication. The investigator global    assessment, IGA, is based upon a rating scale of 0 to 4 with 0=none    or clear, 1=almost clear, 2=mild disease involvement, 3=moderate    disease involvement, and 4=severe disease involvement, and:-   2. separate counts of all types of acne lesions i.e. open and closed    comedones, papules, pustules, nodules, and cysts.

Statistical analyses of each of the efficacy evaluations are performedfor each of the 2 week study time points. Definitive evaluation ofefficacy is based upon comparisons of active to vehicle groups at end oftreatment at 12 weeks. The 4 week-post treatment evaluation is utilizedto evaluate durability of treatment effect after medicationdiscontinuation.

It is to be understood that the foregoing describes preferredembodiments of the present invention and that modifications may be madetherein without departing from the scope of the present invention as setforth in the claims.

What is claimed is:
 1. A method of treating inflammation or pain,comprising the steps of: administering to a subject suffering frominflammation or pain a compound of 3-(methylsulfonyl)acrylonitrile, inan amount effective to treat inflammation or pain.
 2. The methodaccording to claim 1, wherein said method reduces or alleviates thesymptoms of localized manifestations of inflammation characterized byacute or chronic swelling, pain, or redness.
 3. The method according toclaim 1, wherein said inflammation and/or pain is associated with askeletal or muscular disease or condition selected from the groupconsisting of: musculoskeletal sprains, musculoskeletal strains,tendonopathy, peripheral radiculopathy, osteoarthritis, degenerativejoint disease, juvenile arthritis, gout, ankylosing spondylitis,psoriatic arthritis, system lupus erythematosus, costochondritis,tendonitis, bursitis, temporomandibular joint syndrome, andfibromyalgia.
 4. The method according to claim 1, wherein saidinflammation and/or pain is associated with joints, ligaments, tendons,bone, muscles, or fascia.
 5. The method according to claim 1, whereinsaid inflammation and/or pain is associated with an inflammatory skindisease or disorder selected from the group consisting of psoriasis,dermatitis, acne, and rosacea.
 6. The method according to claim 1,wherein said administering is by topical administration.
 7. The methodaccording to claim 1, wherein said administering is by oraladministration.
 8. A method for treating pain, comprising: administeringto the subject suffering from pain an effective amount of morphine andan effective amount of 3-(methylsulfonyl)acrylonitrile.
 9. A method forpotentiating the analgesic effects of morphine, comprising:administering an effective amount of 3-(methylsulfonyl)acrylonitrile toa subject who is being treated with morphine and is suffering from pain.